Pitstop® 2, cell-permeable clathrin inhibitor

• Chemical Structure

Lab

Chemical Structure - Pitstop® 2, cell-permeable clathrin inhibitor (AB120687)

2D chemical structure image of ab120687, Pitstop® 2, Novel cell-permeable clathrin inhibitor

• FuncS

PubMed

Functional Studies - Pitstop® 2, cell-permeable clathrin inhibitor (AB120687)

A) Pitstop® 2 reversibly inhibits Tf uptake. After 15 min preincubation HeLa cells were incubated with Alexa Fluor® 568-Tf in the presence of DMSO or 30 μM Pitstop 2 for 15 min. Tf uptake is seen to resume after washout of the drug for 1 hr. Scale bar, 10 mm. B) Reversibility and dose dependence of Pitstop® 2-mediated inhibition of Tf uptake. Data represent SEM (n = 3 independent experiments; *p < 0.05, ***p < 0.0001). C) Pitstop® 2 inhibits EGF uptake. HeLa cells pretreated with 30 μM pitstop 2 or DMSO for 15 min were incubated for 15 min with Alexa Fluor® 488-EGF in the continued presence of inhibitor. Data represent SEM (n = 3 independent experiments; ***p < 0.0001). D) Pitstop 2 does not interfere with AP-2-mediated cargo sequestration into CCPs. TIRF microscopy images of Cos7 cells pretreated with DMSO or 30 μM Pitstop 2 for 15 min were incubated with Alexa Fluor® 488-EGF at 8oC in the continued presence of inhibitor and immunostained for AP-2a (red). Scale bar, 4 mm. E) Pearson's

Von Kleist L et al., Cell. 2011 Aug 5;146(3):471-84. Fig, 3(A-E)., doi: 10.1016/j.cell.2011.06.025.

• FuncS

PubMed

Functional Studies - Pitstop® 2, cell-permeable clathrin inhibitor (AB120687)

Hela cells were preincubated with DMSO (0.1%) or different doses of pitstop 2 (ab120687) ranging from 5 μM to 30 μM. Cells were then allowed to internalize Alexa594-Transferrin and antibodies to MHCI in the presence or absence of the drug for 30 min. After internalization, surface antibody was removed by low pH acid wash. Cells were then labeled with secondary antibodies to detect trasnferrin and MHCI.

Credit : Dutta D et al. PLoS One. 2012; 7(9) : e45799. doi : 10.1371/journal.pone.0045799

Dutta D et al. PLoS One. 2012; 7(9): e45799. doi: 10.1371/journal.pone.0045799 Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/